Target (protein/gene name): Gyceraldehyde-3-phosphate Dehydrogenase/ GAPDH E.C Number: 1.2.1.12 NCBI Gene # or RefSeq#: AIF58743.1 https://www.ncbi.nlm.nih.gov/protein/AIF58743.1 PDB ID: 4DIB Organism: Bacillus Anthracis Disease Information: Bacillus anthracis is a large, gram positive bacterium that causes anthrax, most commonly in animals and occasionally in humans. Bacillus anthracis is the only pathogen within its genus, bacillus. Anthrax is transmitted from inhaling or ingesting spores in contaminated soil, plants, or water. People get infected with anthrax when the spores get into the body and activate. Once the spores become active, they multiply inside the body and release toxins. To prevent an outbreak in areas with animals that have anthrax, routine vaccines are performed. Although anthrax is very rare in the US, it has been used as a biological weapon in 2001, killing 5 of the 22 that were infected. Bacillus anthracis has a high chance of being used as a biological weapon because of how easy the spores can be found in nature and the spores can be put into foods and water without anyone knowing. https://www.cdc.gov/anthrax/index.html https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1769905/#__sec1title
Essentiality: Glyceraldehyde-3-phosphate dehydrogenase serves as an important enzyme for very diverse functions. GAPDH catalyzes the sixth step of glycolysis, breaking down glucose for energy and carbon molecules. Depending on the location of GAPDH, it has non-glycotic functions. The translocation to the nucleus acts as a signaling mechanism for apoptosis. http://www.ebi.ac.uk/interpro/potm/2004_2/Page3.htm
Target (protein/gene name): Gyceraldehyde-3-phosphate Dehydrogenase/ GAPDH
E.C Number: 1.2.1.12
NCBI Gene # or RefSeq#: AIF58743.1 https://www.ncbi.nlm.nih.gov/protein/AIF58743.1
PDB ID: 4DIB
Organism: Bacillus Anthracis
Disease Information: Bacillus anthracis is a large, gram positive bacterium that causes anthrax, most commonly in animals and occasionally in humans. Bacillus anthracis is the only pathogen within its genus, bacillus. Anthrax is transmitted from inhaling or ingesting spores in contaminated soil, plants, or water. People get infected with anthrax when the spores get into the body and activate. Once the spores become active, they multiply inside the body and release toxins. To prevent an outbreak in areas with animals that have anthrax, routine vaccines are performed. Although anthrax is very rare in the US, it has been used as a biological weapon in 2001, killing 5 of the 22 that were infected. Bacillus anthracis has a high chance of being used as a biological weapon because of how easy the spores can be found in nature and the spores can be put into foods and water without anyone knowing.
https://www.cdc.gov/anthrax/index.html
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1769905/#__sec1title
Essentiality: Glyceraldehyde-3-phosphate dehydrogenase serves as an important enzyme for very diverse functions. GAPDH catalyzes the sixth step of glycolysis, breaking down glucose for energy and carbon molecules. Depending on the location of GAPDH, it has non-glycotic functions. The translocation to the nucleus acts as a signaling mechanism for apoptosis.
http://www.ebi.ac.uk/interpro/potm/2004_2/Page3.htm
Druggable Target: Glyceraldehyde-3-phosphate Dehydrogenase in a different organism, Trypanosoma cruzi, has a druggability of 0.3.
http://www.brenda-enzymes.org/enzyme.php?ecno=1.2.1.12&Suchword=&reference=&UniProtAcc=&organism%5B%5D=Bacillus+anthracis
E.C. # 1.2.1.12
Enzyme Assay:
https://www.biovision.com/documentation/datasheets/K680.pdf
Cost of 3-phospho-D-phosphate: $281.00
Cost of NADH: $130.00
Cost of GAPDH: $61.00
*from Sigma-Aldrich
Structure: 4DIB from PDB
https://www.rcsb.org/structure/4DIB
Current Inhibitors: Known inhibitors 4: R-Deprenyl. HCl, Heptelidic Acid, Deprenyl, CGP 3466 maleate
Expression Information: Has been expressed in E. coli BL21(DE3).
Purification Information: Ammonium sulfate fractionation and one chromatographic step- dye affinity chromatography on Blue Sepharose.
https://file.scirp.org/pdf/ABC_2013043012565131.pdf
PyMol image of Protein:
Fig 1. PyMol structure of Glyceraldehyde-3-phophate dehydrogenase obtained from the PDB. Structure shown in sticks.
Amino Acid Sequence:
AIF58743.1 glyceraldehyde-3-phosphate dehydrogenase [Bacillus anthracis]
MTRVAINGFGRIGRMVFRQAIKESAFEIVAINASYPSETLAHLIKYDTVHGKFDGTVEAFEDHLLVDGKM
IRLLNNRDPKELPWTDLGVEVVIEATGKFNAKEKAILHVEAGAKKVILTAPGKNEDVTIVVGVNEDQLDI
TKHTVISNASCTTNCLAPVVKVLDEQFGIENGLMTTVHAYTNDQKNIDNPHKDLRRARACGQSIIPTTTG
AAKALAKVLPHLNGKLHGMALRVPTPNVSLVDLVVDVKRDVTVEAINDAFKTVANGALKGIVEFSEEPLV
SIDFNTNTHSAIIDGLSTMVMGDRKVKVLAWYDNEWGYSRRVVDLVTLVVDELAKQENVQHI
Length of sequence: 342 amino acids
Molecular Weight using ExPASy: 37.49206 kDa.
Molar Extinction Coefficient at 280 nm: 24075
TMpred graph Image from https://embnet.vital-it.ch/cgi-bin/TMPRED_form_parser:
Blast:
Query Coverage: 100%
Max % Identities: 100%
% Positives: 100%
Chain used for Homology: A
CDS Gene Sequence (codon optimized): **
ATGACCCGCGTGGCGATTAACGGCTTTGGCCGCATTGGCCGCATGGTGTTTCGCCAGGCG
ATTAAAGAAAGCGCGTTTGAAATTGTGGCGATTAACGCGAGCTATCCGAGCGAAACCCTG
GCGCATCTGATTAAATATGATACCGTGCATGGCAAATTTGATGGCACCGTGGAAGCGTTT
GAAGATCATCTGCTGGTGGATGGCAAAATGATTCGCCTGCTGAACAACCGCGATCCGAAA
GAACTGCCGTGGACCGATCTGGGCGTGGAAGTGGTGATTGAAGCGACCGGCAAATTTAAC
GCGAAAGAAAAAGCGATTCTGCATGTGGAAGCGGGCGCGAAAAAAGTGATTCTGACCGCG
CCGGGCAAAAACGAAGATGTGACCATTGTGGTGGGCGTGAACGAAGATCAGCTGGATATT
ACCAAACATACCGTGATTAGCAACGCGAGCTGCACCACCAACTGCCTGGCGCCGGTGGTG
AAAGTGCTGGATGAACAGTTTGGCATTGAAAACGGCCTGATGACCACCGTGCATGCGTAT
ACCAACGATCAGAAAAACATTGATAACCCGCATAAAGATCTGCGCCGCGCGCGCGCGTGC
GGCCAGAGCATTATTCCGACCACCACCGGCGCGGCGAAAGCGCTGGCGAAAGTGCTGCCG
CATCTGAACGGCAAACTGCATGGCATGGCGCTGCGCGTGCCGACCCCGAACGTGAGCCTG
GTGGATCTGGTGGTGGATGTGAAACGCGATGTGACCGTGGAAGCGATTAACGATGCGTTT
AAAACCGTGGCGAACGGCGCGCTGAAAGGCATTGTGGAATTTAGCGAAGAACCGCTGGTG
AGCATTGATTTTAACACCAACACCCATAGCGCGATTATTGATGGCCTGAGCACCATGGTG
ATGGGCGATCGCAAAGTGAAAGTGCTGGCGTGGTATGATAACGAATGGGGCTATAGCCGC
CGCGTGGTGGATCTGGTGACCCTGGTGGTGGATGAACTGGCGAAACAGGAAAACGTGCAG
CATATT